Department of Obstetrics and Gynaecology, National Institute for Health Research Cambridge Biomedical Research Centre, University of Cambridge School of Clinical Medicine; Centre for Trophoblast Research, University of Cambridge.
Department of Obstetrics and Gynaecology, National Institute for Health Research Cambridge Biomedical Research Centre, University of Cambridge School of Clinical Medicine.
J Vis Exp. 2021 Oct 14(176). doi: 10.3791/62670.
Described here is a simple method to isolate and phenotype mouse group 1 uterine innate lymphoid cells (g1 uILCs) from individual pregnant uterus by flow cytometry. The protocol describes how to set up time mating to obtain multiple synchronous dams, the mechanical and enzymatic digestion of the pregnant uterus, the staining of single-cell suspensions, and a FACS strategy to phenotype and discriminate g1 uILCs. Although this method inevitably loses the spatial information of cellular distribution within the tissue, the protocol has been successfully applied to determine uILC heterogeneity, their response to maternal and foetal factors affecting pregnancy, their gene expression profile, and their functions.
本文介绍了一种通过流式细胞术从单个妊娠子宫中分离和表型鉴定小鼠第 1 组固有淋巴样细胞（g1 uILCs）的简单方法。该方案描述了如何设置时间交配以获得多个同步的孕鼠，对妊娠子宫进行机械和酶消化，单细胞悬液的染色，以及一种 FACS 策略来表型鉴定和区分 g1 uILCs。尽管该方法不可避免地丢失了细胞在组织内分布的空间信息，但该方案已成功应用于确定 uILC 的异质性、它们对影响妊娠的母体和胎儿因素的反应、它们的基因表达谱以及它们的功能。
